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Nucleic Acids Research, 1990, Vol. 18, No. 13 3739-3744
© 1990

MOLECULAR BIOLOGY

High fidelity DNA synthesis by the Thermus aquaticus DNA polymerase

Kristin A. Eckert and Thomas A. Kunkel*

Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences PO Box 12233, Research Triangle Park, NC 27709, USA

* To whom correspondence should be addressed

Received April 18, 1990. Revised May 22, 1990. Accepted May 22, 1990.

We demonstrate that despite lacking a 3' — 5' proofreading exonuclease, the Thermus aquaticus (Taq) DNA polymerase can catalyze highly accurate DNA synthesis in vitro. Under defined reaction conditions, the error rate per nucleotide polymerized at 70°C can be as low as 10–5 for base substitution errors and 10–6 for frameshift errors. The frequency of mutations produced during a single round of DNA synthesis of the lac Z{alpha} gene by Taq polymerase responds to changes in dNTP concentration, pH, and the concentration of MgCI2 relative to the total concentration of deoxynucleotide triphosphates present in the reaction. Both base substitution and frameshift error rates of < 1/100, 000 were observed at pH 5–6 (70°C) or when MgCI2 and deoxynucleotide triphosphates were present at equimolar concentrations. These high fidelity reaction conditions for DNA synthesis by the Taq polymerase may be useful for specialized uses of DNA amplified by the polymerase chain reaction.


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